BACKGROUND: During blood bank storage, red blood cells (RBCs) undergo a number of biological and biochemical alterations collectively referred to as "storage lesions". These injuries include loss and oxidative cross-linking of band 3, the major integral protein of RBC membranes. Denaturation of hemoglobin (Hb) and damage to the amino-terminal of band 3 are recognised as the starting events for immunological recognition mechanisms and phagocytic removal of senescent or impaired RBCs from circulation. Consequently, studies focusing on the Hb-association and oxidative status of the cytoskeleton of stored RBCs intended for transfusion are of extreme interest. In this work, two storage-related fragments of band 3 were documented and biochemically characterised. METHODS: Four RBC units were collected from normal volunteers and stored for 21 days under (i) standard blood bank conditions, (ii) anaerobic conditions, or (iii) in the presence of caspase 3-inhibitor. Degradation products of band 3 were followed by sodium dodecyl sulfatepolyacrylamide gel electrophoresis coupled with western blot and mass spectrometry analyses. RESULTS: Two different degradation products of the cytoplasmic domain of the erythrocyte band 3 (CDB3) were detected in RBC membranes during storage in saline-adenine-glucosemannitol (SAGM) preservation medium. One of these fragments showed an apparent molecular weight of 34 kDa and was demonstrated to be the product of a free-radical attack on the protein main chain, whereas another fragment of 24 kDa was the result of a caspase 3-mediated cleavage. DISCUSSION: Although to different extent, anaerobic conditions reduced the formation of both truncated products indicating an enhanced activity of the pro-apoptotic caspase 3 enzyme following oxidative stress. Interestingly, both CDB3 fragments were tightly associated to the erythrocyte membrane supporting the involvement of Cys-201 and/or Cys-317 in clustering different band 3 monomers.
BACKGROUND: During blood bank storage, red blood cells (RBCs) undergo a number of biological and biochemical alterations collectively referred to as "storage lesions". These injuries include loss and oxidative cross-linking of band 3, the major integral protein of RBC membranes. Denaturation of hemoglobin (Hb) and damage to the amino-terminal of band 3 are recognised as the starting events for immunological recognition mechanisms and phagocytic removal of senescent or impaired RBCs from circulation. Consequently, studies focusing on the Hb-association and oxidative status of the cytoskeleton of stored RBCs intended for transfusion are of extreme interest. In this work, two storage-related fragments of band 3 were documented and biochemically characterised. METHODS: Four RBC units were collected from normal volunteers and stored for 21 days under (i) standard blood bank conditions, (ii) anaerobic conditions, or (iii) in the presence of caspase 3-inhibitor. Degradation products of band 3 were followed by sodium dodecyl sulfatepolyacrylamide gel electrophoresis coupled with western blot and mass spectrometry analyses. RESULTS: Two different degradation products of the cytoplasmic domain of the erythrocyte band 3 (CDB3) were detected in RBC membranes during storage in saline-adenine-glucosemannitol (SAGM) preservation medium. One of these fragments showed an apparent molecular weight of 34 kDa and was demonstrated to be the product of a free-radical attack on the protein main chain, whereas another fragment of 24 kDa was the result of a caspase 3-mediated cleavage. DISCUSSION: Although to different extent, anaerobic conditions reduced the formation of both truncated products indicating an enhanced activity of the pro-apoptotic caspase 3 enzyme following oxidative stress. Interestingly, both CDB3 fragments were tightly associated to the erythrocyte membrane supporting the involvement of Cys-201 and/or Cys-317 in clustering different band 3 monomers.
Authors: J A Walder; R Chatterjee; T L Steck; P S Low; G F Musso; E T Kaiser; P H Rogers; A Arnone Journal: J Biol Chem Date: 1984-08-25 Impact factor: 5.157
Authors: M M Kay; S R Goodman; K Sorensen; C F Whitfield; P Wong; L Zaki; V Rudloff Journal: Proc Natl Acad Sci U S A Date: 1983-03 Impact factor: 11.205
Authors: Julie A Reisz; Travis Nemkov; Monika Dzieciatkowska; Rachel Culp-Hill; Davide Stefanoni; Ryan C Hill; Tatsuro Yoshida; Andrew Dunham; Tamir Kanias; Larry J Dumont; Michael Busch; Elan Z Eisenmesser; James C Zimring; Kirk C Hansen; Angelo D'Alessandro Journal: Transfusion Date: 2018-10-12 Impact factor: 3.157
Authors: Walter H Reinhart; Nathaniel Z Piety; Jeremy W Deuel; Asya Makhro; Thomas Schulzki; Nikolay Bogdanov; Jeroen S Goede; Anna Bogdanova; Rajaa Abidi; Sergey S Shevkoplyas Journal: Transfusion Date: 2015-03-06 Impact factor: 3.157